| 表达猪白细胞介素2重组腺病毒的构建 |
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| 引用本文： | 张健,陈艳,乔传玲,杨焕良,唐续,辛晓光,陈化兰.表达猪白细胞介素2重组腺病毒的构建[J].中国预防兽医学报,2011,33(4). |
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| 作者姓名： | 张健 陈艳 乔传玲 杨焕良 唐续 辛晓光 陈化兰 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部动物流感重点开放实验室,黑龙江哈尔滨,150001 |
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| 摘 要： | 为构建表达猪白细胞介素2(IL-2)的复制缺陷型重组腺病毒,本研究提取经刀豆球蛋白A(con A)刺激体外培养的长白猪外周血淋巴细胞总RNA,利用RT-PCR扩增猪IL-2基因,并克隆于质粒pIRES2-EGFP中,将含有IL-2及EGFP的基因片段亚克隆到腺病毒的穿梭质粒pDC315中,构建重组腺病毒穿梭质粒pDC315-IL-2-EGFP.利用脂质体转染方法将pDC315-IL-2-EGFP和腺病毒骨架质粒pBHGloxΔEl,E3Cre共转染HEK293细胞.根据腺病毒感染后形成的典型细胞病变及EGFP的荧光信号筛选重组腺病毒rAd-IL-2-EGFP.经western blot鉴定,结果表明获得了一株能够表达猪IL-2蛋白的重组腺病毒rAd-IL-2-EGFP.本研究为进一步研究猪IL-2在疫苗免疫中的增强作用及研制新型免疫增强剂奠定了基础.
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| 关 键 词： | 重组腺病毒 基因表达 |
Construction of recombinant adenovirus expressing porcine interleukin 2 gene |
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| ZHANG Jian,CHEN Yan,QIAO Chuan-ling,YANG Huan-liang,TANG Xu,XIN Xiao-guang,CHEN Hua-lan.Construction of recombinant adenovirus expressing porcine interleukin 2 gene[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(4). |
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| Authors: | ZHANG Jian CHEN Yan QIAO Chuan-ling YANG Huan-liang TANG Xu XIN Xiao-guang CHEN Hua-lan |
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| Institution: | ZHANG Jian,CHEN Yan,QIAO Chuan-ling*,YANG Huan-liang,TANG Xu,XIN Xiao-guang,CHEN Hua-lan (Animal Influenza Laboratory of the Ministry of Agricultural,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
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| Abstract: | To construct a recombinant adenovirus expressing porcine interleukin 2 (IL-2), The IL-2 gene was amplified by RT-PCR with specific primers from the total RNA, which were extracted from peripheral blood lymphocytes after stimulation with ConA in vitro and cloned into pIRES2-EGFP. The recombinant adenovirus shuttle plasmid pDC315-IL2-EGFP was constructed by inserting IL2-IRES-EGFP into shuttle plasmid pDC315. A recombinant adenovirus expressing IL-2 (rAd-IL-2-EGFP) was generated by co-transfecting the pDC315-... |
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| Keywords: | IL-2 |
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